GLUCOSIDES  OF  EMODIN 


BY 


LEONARD  ALBERT  STIDLEY 

A.  B.  Carthage  College,  1921 


THESIS 

SUBMITTED  IN  PARTIAL  FULFILLMENT  OF  THE  REQUIREMENTS 
FOR  THE  DEGREE  OF  MASTER  OF  SCIENCE  IN  CHEMISTRY 
IN  THE  GRADUATE  SCHOOL  OF  THE  UNIVERSITY 
OF  ILLINOIS,  1922 


URBANA,  ILLINOIS 


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UNIVERSITY  OF  ILLINOIS 


THE  GRADUATE  SCHOOL 


4^ 19^ 


I HEREBY  RECOMMEND  THAT  THE  THESIS  PREPARED  UNDER  MY 

SUPERVISION  BY Leonair.,  .,  ..L_  -r..  J 

ENTITLED  _ - giucosiaes EJlQClin 


BE  ACCEPTED  AS  FULFILLING  THIS  PART  OF  THE  REQUIREMENTS  FOR 


Recommendation  concurred  in* 


Committee 


on 


Final  Examination* 


^Required  for  doctor’s  degree  but  not  for  master’s 


Digitized  by  the  Internet  Archive 
in  2015 


https://archive.org/details/glucosidesofemodOOstid 


ACKNO’ii/LEDGUiENT 

The  writer  wishes  to  express  his  sincdre 
appreciation  for  the  helpful  suggestions  and 
encouragement  from  Dr..  George  D.  Beal,  under 
whose  supervision  this  investigation  was  carried 
out. 


- 1 - 
I 

IlITRODUCTION  AITD  STATEMENT  OF  PROBLEM 

The  name  glucoside  is  applied  to  those  substances,  which 
under  proper  conditions  furnish  when  hydrolysed  by  an  enzyme  or 
an  acid,  a sugar,  usually  dextro-glucose  and  one  or  more  other 
compounds.  Representatives  of  nearly  every  class  of  organic  com- 
pounds occur  in  plants,  chiefly  in  the  fruit,  bark  and  roots, 
in  combination  with  a sugar. 

These  natural  glucosides  first  drew  attention  in  plant  anal- 
ysis, because  of  their  various  useful  properties  in  medicine  and 
the  arts.  The  properties  of  the  hydrolyised  constituents  have 
been  studied  more  than  the  glucosides  themselves. 

Some  of  the  naturally  occurring  glucosides  have  been  prepared 
synthetically.  These  syntheses  have  included  the  classes  of  alco- 
hols, phenols,  aldehydes,  ketones,  alkyls,  amines,  mercaptans, 
and  acids  combined  with  sugar,  Goyre spending  representatives  of 
these  classes  have  been  found  to  occur  in  plants.  Investigations 
of  plants  have  revealed  natural  glucosides  of  anthraqui nones  and 
sugars.  This  work  was  undertaken  with  the  hope  of  synthesizing  a 
dihydroxy  anthraquinone  and  trihydroxymethylanthraquinone-glucoside,. 
corresponding  to  the  form  that  is  found  in  the  plants. 


- 3 - 
II 

HISTORICAL 

The  hydroxyanthraquinones  have  long  been  known  empirically, 
because  of  their  dyeing  and  purgative  properties.  Before  the 
synthesis  of  dyes,  alizarin  was  the  most  valuable  natural  material 
in  the  dyeing  industry. 

Alizarin,  1-3  dihydroxyanthrquinone,  is  the  principle  con- 
stituent of  the  madder  root,  Rubia  tinctorium,  in  which  it  occurs 

as  a glucoside,  ruberythric  acid.  The  first  discovery  of  import- 

(1) 

anoe  was  made  by  Robiquet  and  Colin,  who  in  1836,  succeeded  in 
preparing  from  madder  a substance  which  on  being  subjected  to  heat 
gave  a sublimate  consisting  of  beautiful  reddish-yellow  needles, 
and  to  which  they  gave  the  name  alizarin.  The  method  which  they 
adopted  left  it  quite  uncertain  whether  or  not  it  was  a product 
of  decomposition  of  some  other  body  formed  by  the  action  of  heat. 
The  same  series  of  experiments  led  to  the  discovery  of  another 
coloring  mtter  in  the  madder  to  which  these  chemists  gave  the 
namie  purpurine.  This  was  also  a hydroxyanthraquinone  glucoside. 

In  the  year  of  1835  a memoir  was  published  by  Runge  containing 
a deLscription  of  the  properties  and  methods  of  oreparation  of 

(3) 

three  distinct  coloring  matters  from  madder.  In  1838  Decaisne 
in  an  extensive  study  of  the  madder  root  stated  that  the  effect 

of  oxygen  upon  the  yellow  colored  substance  produced  the  red  dye, 
(3) 

Rochleder  had  the  view  first  that  alizarin  and  the  glucose  were 
connected  in  ruberythric  acid  in  the  ratio  of  two  molecules  of 


- 3 - 

glucose  to  one  of  alizarin. 

^ HgO  ^ 3 C0H13O6  C14K8O4 

Ruberythric  acid  was  obtained  from  extracting  powdered  madder 
root  with  absolute  alcohol.  The  yield  was  0.1^  of  the  v/eight 
of  the  dried  root.  The  compound  formed  lemon-colored  needles 
of  silky  luster,  which  melted  at  358-860  degrees.  In  1860, 

(4) 

Schunk  claimed  to  have  isolated  another  glucoside  which  yielded 

an  anthraquinone  upon  hydrolysis.  This  new  substance  he  called 

rubianic  acid,  and  which  he  did  not  think  was  identical  with 

Rochleder' s ruberythric  acid.  Schunk  claimed  that  it  did  not 

pre-exist  in  the  plant,  but  that  it  v/as  formed  by  the  action  of 

oxygen  upon  a more  complex  body.  Schunk  showed  that  alizarin 

and  purpurine  T;ere  allied  substances,  since  both  yielded  phthalic 

acid  when  they  were  decomposed  by  nitric  acid,  and  napthalene  was 

(5) 

the  only  substance  which  was  known  to  yield  this.  Stokes  drew 

up  a table  to  show  the  differences  in  physical  properties  between 

(6) 

alizarin  and  purpurine.  Schunk  and  Ivlarchlewski  showed  that  rubi-^ 

j 

anic  acid  was  a methyl  substitution  product  of  purpuroxanthin, 
which  was  an  isomer  of  alizarin,  obtained  from  purpurine  by  "de- 
oxidation. ” They  also  isolated  another  hydroxyanthraquinone 
glucoside  (rubiadin  glucoside)  which  yielded  upon  hydrolysis, 
rubj^in  and  glucose.  Liebermann  and  Bergami  showed  that  rubery- 
thric acid  contained  one  of  the  two  hydroxy  groups  of  alizarin 
in  a free  state,  and  that  the  union  of  the  alizarin  with  the 

sugar  took  place  through  that  hydroxyl  which  may  be  easily 


- 4 - 

alkylated.  Since  ruberythrio  acid  yields  am  octa-acetyl  deriv- 
ative the  following  formulae  for  the  compound  were  suggested  with 
preference  to  the  second: 


(1)  g,,h.o 


^ V 

- \oC5H^^03 


(2) 


14^6 


H^O 


00^2^2 1®10 
OH 


(8) 


In  1897  Perkin  studied  the  yellow  coloring  matter  of  the  madder 

(9) 

root,  but  isolated  no  new  products.  LJuller  and  de  la  P.ue 

isolated  from  rhubarb  root  some  anthraquinone  glucosides,  which 

yielded  emodin  and  chrysophanic  acid  upon  hydrolysis.  Later, 

(10) 

Muller  isolated  alizarin  by  hydrolysis  of  the  same  root.  Per- 
haps no  drug  recognized  by  any  of  the  national  pharmacopoeias 

has  more  frequently  engaged  the  attention  of  chemists  than  rhubarb. 

(11) 

According  to  Tutin  and  Clewer  , the  anthraquinone  glucosides 

which  have  been  isolated  from  that  plant  are  rhein,  emodin,  aloe- 

emodin,  emodin  monomethyl  ether,  chrysophanic  acid  and  rhelnolic 

(13) 

aold.  Jowett  and  Potter  analvsed  araroba  to  study  ohrysarotln, 

(13) 

the  anthranol  of  chrysophanic  acid.  Tutin  found  that  anthra- 
quinone derivatives  present  in  senna  were  rhein  and  aloe-emodin. 
(14) 

Bailey  considers  that  chrysophanic  acid  is  a constituent  of 
senna.  The  bark  and  fruit  of  several  species  of  the  Rhamonaoaae 
contain  several  anthraquiijjes  in  combination  about  which  there  is 
some  dispute.  The  buckthorns  contain  a glucoside  to  which  the 
name  frangulin  has  been  given  and  which  yields  upon  hydrolysis 


- 5 - 


rhamnose  and  emodln. 


li  0 

? 1 9 ? 


G H , 0^-f-  G H 0^ 
6 14  6 15  10  5 

(15) 


Frangulin  ms  first  separated  by  Blns'.vanger  in  Rharmus  francmla, 

(16) 

and  ms  recognized  as  a glucoside  by  Faust.  Frangulinic  acid 

is  the  primary  glucoside  of  Rhamnus  frangula  and  has  been  iso- 

(17)'  (18) 


lated  in  Cascara  segrada. 


Gunton  in  his  work 


on  a rein- 


vdstlgation  of  the  proximate  composition  of  Rhamnus  frangula  en- 
countered the  natural  frangulin,  and  subsequently  7/orked  upon  the 
synthesis  of  the  glucosides  of  this  type, 

(19) 

The  s^mthetic  ’work  on  glucosides  was  begun  then  Colley 
by  the  use  of  acetocloroglucose  was  able  to  effect  a combination 


of  this  compound  with  the  potassi'um  salt  of  the  phenols  and  alco- 
(30) 

hols.  Micheal  was  able  to  combine  glucose  in  1he  form  of  the 

acetoohloroglucose  with  a simple  phenol.  This  product  agreed  ’with 

the  natural  o courting  glucoside.  This  was  followed  by  the  syn- 

(31) 

thesis  of  helicin,  salicin,  and  methyl  arbutin.  Plugo  Schiff 

produced  compounds  of  aldehydes  and  ketones  with  the  sugars  thru 

the  union  of  the  components  in  acetic  acid  solution.  By  means  of 

the  acetoohloroglucose  and  the  corresoonding  bromo  derivative, 

(33) 

Bruin  formed  the  thymol glucoside  and  the  ©<  -napthol  glucoside, 

, , (33)  (34)1 

Irvine  synthesized  aaiino  glucosides  from  d-glucosamine.  Ryan 

(35) 

used  arablnose  and  xylose  to  form  the  glucosides.  Llauthner 

(36f 

synthesized  acid  glucosides,  Salaay  fatty  alcohol  glucosides 
(37)  (38) 

and  Schneider  mustard  oil  glucosides.  Fischer  and  his 


students  have  synthesized  the  alkyl  and  phenolic  glucosides. 


6 


(1§) 

Gunton  has  been  the  only  one  to  work  upon  the  synthesis  of 
the  anthraquinone  gluco sides. 


- 7 - 
III 

PREPARATION  OF  MATERIAL. 

(39) 

I.  Preparation  of  Rhamnose. 

Following  the  method  of  Walton,  two  hundred  grams  of  lemon 
flavin  were  suspended  in  two  liters  of  water,  acidified  with 
about  0.5^  sulphuric  acid  and  boiled  for  three  hours.  The  yellow 
plastic  material  was  filtered  off  and  the  filtrate  neutralized 
with  barium  carbonate.  It  was  then  clarified  with  bone  charcoal 
which  removed  the  creamy  color.  The  filtrate  was  then  concen- 
trated under  diminished  pressure  to  a thick  syrup  of  about  40fo 
solids.  This  sugar  was  diluted  to  three  times  its  volume  of  abso- 
lute alcohol  and  the  rhamnose  crystallized  out  by  concentrating 
the  filtrate  to  a density  of  70-60^  solids.  Pure  white  crystals 
of  rhamnose  were  obtained  after  recrystallization  from  absolute 
alcohol.  The  osazone  melted  at  180°.  The  first  preparation 
yielded  18^  of  the  weight  of  the  lemon  flavin  taken,  and  the 
second  yielded  30^. 

(30) 

II,  Preparation  of  Alizarin, 

Indicator  Alizarin  was  used.  It  was  sublimed  by  placing  a 
beaker  through  which  cold  water  was  circulating  over  a crucible 
containing  the  alizarin.  A Bunsen  burner  was  applied  gently  until 
the  sublimate  could  be  seen  forming.  The  flame  ms  kept  at  that 
heat.  The  yield  was  so  small  that  it  was  necessary  to  synthesize 
some  alizarin. 


8 


Ten  grants  of  anthraquinone  and  85  cc.  of  fuining  sulphuric 
acid  vfsre  placed  in  a small  flask  and  heated  to  300-230®C.  on  an 
oil -bath,  raising  the  temperature  slowly,  for  about  two  hours. 

The  mixture  was  allowed  to  cool  and  poured  into  300  cc,  of  water, 
then  filtered.  50  grams  of  salt  was  then  added,  and  the  solution 
stirred  thoroughly  and  allowed  to  stand  in  cold  water  until  the 
sodium  anthraquinone  sulphonate  separated.  The  salt  was  filtered 
off,  pressed  and  dried  on  a porous  plate.  Into  an  iron  pipe  which 
could  be  closed  at  both  ends  by  means  of  a screw  cap  were  put 
40  grams  of  sodium  hydroxide,  10  grams  of  sodium  anthraquinone- 
sulphonate,  3 grams  of  potassium  chlorate  and  40  cc.  of  water. 

The  bomb  v/as  heated  in  an  electric  oven  for  30  hours  at  170®C. 

The  dark  blue  mass  which  resulted  was  dissolved  in  hot  water, 
filtered  and  neutralized  v;ith  hydrochloric  acid.  The  alizarin 
which  was  then  precipitated  was  filtered  off  and  dried.  It  v/as 
recrystallized  from  glacial  acetic  acid. 

(31) 

Preparation  of  Tetra-acetyl-bromo-glucose. 

The  method  of  Dale  was  used  with  several  minor  alterations, 
T'.vanty-f ive  grams  of  glucose  were  shaken  in  135  cc.  of  acetic  an- 
hydride. liydrobromic  acid  from  the  action  of  bromine  upon  red 
phosphorous  was  bubbled  through  the  suspension  until  the  acetic 
anhydride  v;as  saturated.  The  solution  became  a light  green, 
and  the  glucose  rapidly  dissolved  with  the  evolution  of  consider- 
able heat.  After  cooling  the  solution  was  dissolved  in  300  cc. 


- 9 - 

of  chloroform  and  the  resulting  solution  washed  to  remove  the 
acid,  once  with  water  and  once  with  a solution  of  sodium  bicarbon- 
ate, and  then  dried  with  calcium  chloride  to  remove  the  water. 

It  was  then  evaporated  under  diminished  pressure  to  a thick 
yellow  syrup,  washed  into  a beaker  with  a small  quantity  of  ether 
and  the  tetra-acetyl-bromo-glucose  was  precipitated  with  15  vol- 
umes of  petroleum  ether.  The  compound  came  out  in  a semi-liquid 
state  which  solidified  upon  stirring.  It  had  a yellow  color  and 
an  acetyl  odor.  After  recrystalizing  twice  by  dissovling  in  75 
cc.  of  ether  and  evaporating  the  solution  in  a current  of  dry 
air  until  crystals  began  to  appear  the  odor  was  lost  and  the  com- 
pound was  White.  A sample  melted  at  88"  agreeing  with  that  of 
Dale,  The  various  quantities  which  were  made  varied  in  yields 
from  65 -7 0'^ 

Preparation  of  aoetobromo -rhamnose 
Thirty  grams  of  rhamnoae  were  suspended  in  one  hundred  and 
fifty  cc.  of  acetic  anhydride  and  hydrobromic  acid  prepared  as 
in  the  case  of  acetobromoglucose,  passed  into  the  solution  until 
it  was  saturated.  The  solution  became  a light  brovTn  and  the 
rhamnose  dissolved  with  the  evolution  of  considerable  heat.  This 

solution  was  treated  similarly  to  one  in  the  above  mentioned 
preparation.  The  product  obtained  was  a dark  brown  syrup  having 
an  acetyl  odor.  Attempts  were  made  to  crystalize  it  from  several 
solvents,  but  it  still  remained  a liquid.  The  total  yield  was 
forty  grams.  The  compound  kept  v/ell  in  stoppered  bottles. 


- 10  - 
A 

Alizarin  Glucoside. 

Five  grams  of  acetylbromoglucoss  were  added  to  an  absolute 
alcohol  solution  of  3.66  grams  of  alizarin  and  1.'33  grams  of 
potassium  hydroxide  in  an  Erlenmeyer  flask.  The  flask  was  tightly 
stoppered  and  on  shaking  the  mixture  became  a deep  violet  due  to 
the  formation  of  the  potassium  salt  of  the  alizarin.  This  mixture 
was  allowed  to  stand  at  outside  temperature  for  one  week  with 
occasional  shaking.  It  was  then  refluxed  for  one  and  one  half 
hours.  There  was  no  visible  change  in  the  mixture  at  this  time, 
for  the  deep  violet  color  persisted.  The  alcoholic  solution  was 
then  diluted  with  water  and  exactly  neutralized  with  hydrochloric 
acid,  which  precipitated  the  dissolved  material  as  a fine  orange 
precipitate,  having  a strong  odor.  This  was  filtered  off,  dried 
and  weighed.  It  weighed  3.67  grams.  The  orange  bro^^Ti  precipitate 
showed  clearly  that  a mixture  was  present.  No  melting  point  could 
be  secured.  Part  of  the  substance  was  free  alizarin  which  had 
not  entered  into  the  reaction.  The  precipitate  gave  tests  for 
the  presence  of  glucose. 

Part  of  the  precipitate  was  hydrolysed  with  dilute  sulphuric 
acid.  The  solution  turned  a yellow  shade,  crystals  of  alizarin 
were  evident,  and  the  solution  gave  test  for  the  presence  of  glu- 
cose. It  was  thought  that  this  might  have  been  present  at  the 
beginning  of  the  hydrolysis,  so  the  precipitate  was  washed  and 
extracted  with  ether.  The  ether  v;as  evaporated  and  the  residue 
was  alizarin  as  was  shovm  by  the  orange  crystals  and  the  resulting 
melting  point. 


- 11 


The  remaining  precipitate  was  washed  virith  water  until  the 
wash  water  gave  no  tests  for  the  presence  of  a carbohydrate. 

Then  a portion  was  hydrolysed  with  dilute  sulphuric  acid.  The 
resulting  solution  gave  a test  for  sugar.  It  is  possible  that 
this  may  have  resulted  from  an  occlusion,  rather  than  the  forma- 
tion of  a compound.  Benzene,  methyl  alcohol,  ethyl  alcohol, 
arnyl  alcohol,  and  chloroform  were  used  as  solvents  in  trying  to 
secure  a crystal ine  product,  but  no  positive  results  were  obtained, 
The  melting  point  at  about  83®C.  was  not  sharp.  The  precipitate 
was  sorted  mechanically,  and  vdiat  appeared  to  be  the  best  portion 
as  taken  and  hydrolysed  with  dilute  sulphuric  acid,  but  the 
resulting  solution  did  not  give  tests  for  the  presence  of  glucose. 

A check  upon  the  experiment  failed  to  y’eLd  any  other  product, 
of  a glucosidic  nature. 

Owing  to  the  fact  that  there  was  an  excess  of  alizarin, in 
the  first  experiment,  another  experiment  tise  same  as  the  first, 
but  the  amount  of  alizarin  was  reduced  to  2.5  grams.  After  re- 
fluxing for  one-half  hour  the  solution  was  diluted  and  exactly 
neutralized  with  hydrochloric  acid.  The  same  orange  colored 
precipitate  was  given.  This  was  filtered  and  dried.  ITo  definite 
melting  point  was  obtained.  It  wasLthen  washed  with  ether  to 
remove  the  free  alizarin.  The  ether  was  evaporated  and  some  free 
alizarin  recovered  as  was  shown  by  the  melting  point.  The  original 
yellov;  precipitate  showed  no  evidences  of  cr^rstalline  form. 


1 1 


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- 13 


Alcohol,  acetone  and  chloroform  v;ere  used  in  an  effort  to 
crystallize  the  powder,  but  because  of  its  insolubility,  nothing 
crystalline  could  be  obtained.  The  substance  gave  tests  for  the 
presence  of  carbohydrates  both  before  and  after  hydrolysis.  The 
remaining  precipitate  was  washed  with  water  until  no  tests  for 
sugar  were  given.  Upon  hydrolysis  a slight  test  was  obtained. 
V/hether  or  not  this  was  due  to  an  occlusion  or  to  a combination 
could  not  be  certain,  hut  because  of  the  evident  mixture  of  the 
substance  the  former  seems  more  plausible. 

It  was  thought  that  perhaps  a catalyst  might  aid  in  the 
reaction  so  the  same  proce.dure  as  in  the  last  experiment  was 
carried  out  with  the  addition  of  some  mercury.  The  results 
failed  to  yield  any  evidence  other  than  what  v/ere  given  by  the 
previous  experiments. 

Thinking  that  perhaps  the  chlorine  atom  might  be  mors  easily 
replaced  on  the  acetyl  derivative,  5 grams  of  acetyl  cl^loro glucose 
prep3.red  by  the  same  method  as  the  acetyl bromo  glucose,  v/ere 
added  to  an  Erisnmeyer  flask  containing  an  absolute  alcohol 
solution  of  2.00  grams  of  alizarin  and  1.33  grams  of  potassium 
hydroxide.  This  was  refluxeddfor  one  hour  and  a half,  diluted 

and  exadstly  neutralized  with  hydroohlorio  acid.  A light  orange 
precipitate  resulted.  This  wa.s  filtered,  washed  and  dried.  It 
proved  to  be  a mixture,  and  nothing  crystalline  could  be  isolated 


from  it. 


: VJ:  '.  ’; 


:v*  '■rr.-^r ..  r-  -v  ..  . „ 


- 13 


(33) 

The  successfiul  synthesis  of  recorcinal  o<, -glue  ©side  by 
the  shaking  process  tempted  us  to  try  a similar  procedure,  because 
of  the  similarity  between  recorcinol  and  alizarin.  Five  grams 
of  acetobromoglucose , 2.86  graiiis  of  alizarin  and  1.33  grams  of 
potassium  hydroxide  were  added  to  200  cc.  of  water.  A blue  solu- 
tion resulted.  This  sDolution  was  added  to  a bottle  containing 
400  cc,  of  ether.  The  bottle  was  securely  attached  to  cross-arm 
support  which  was  in  turn  connacted  to  a shaking  machine  driven 
by  a motor.  The  bottle  was  shaken  intermittently  for  tv/enty-four 
hours.  200  cc.  of  ether  were  then  added  and  the  bottle  shaken 
for  another  tv/enty-f our  hours,  A heavy  foam  had  appeared  in  the 
ethereal  layer.  This  layer  was  separated  from  the  aquous  solu- 
tion, and  the  ether  evaporated.  A blue  condensation  product 
resulted.  This  substance  proved  to  be  insoluble  in  the  ordinary 
organic  solvents.  A portion  was  subject  to  hydrolysis  but  did 
not  yeild  any  carbohydrate  test  even  after  it  had  been  hydrolysed 
for  six  hours.  The  product  did  not  give  a melting  point  below 
300  degrees,  nothing  crystalline  could  be  isolated  from  the 
blue Ish -black  powder. 

(33) 

A recent  modification  for  phenolic  glucosides  by  Fischer 
was  tried  with  the  hydroxyanthraquinone.  Five  grams  of  acetobromo- 
glucose and  2.86  grams  of  alizarin  were  intimately  mixed  in  a 
small  beaker,  to  which  was  added  four  grams  of  dried  quinoline. 

The  beaker  was  then  heated  upon  a water  bath  for  two  hours.  It 
was  then  cooled  through  shaking  ’with  100  cc.  N Sulphuric  acid  and 
75  cc.  of  chloroform,  not  all  of  the  substance  dissolved.  The 


- 14  - 


o-iloroform  was  filtersd  after  it  had  teen  washed  with  water  sever- 
al times.  The 3 chloroform  solution  was  then  evaporated  upon  a 
steam  bath.  A light  brov/n  viscous  liquid  with  some  uncombined 
alizarin  remained. 

For  the  conversion  into  the  acetate  product,  a portion  of 
the  viscouassubstance  was  mixed  with  5 cc.  of  dried  pyridine  and 
10  cc.  of  acetic  anhydride.  After  slight  shaking  this  dissolved. 
After  standing  34  hours  this  solution  was  poured  into  ice  water, 
and  the  light  broi^m  liquid  collected  at  the  bottom. 

To  replace  the  acetyl  groups  by  hydroxyls,  two  gfams  of 
the  tetra-acetyl  product  were  dissolved  in  100  cc.  of  v;ater  which 
contained  8 grams  of  crystallized  barium  hydroxide,  and  35  cc. 
of  alcohol.  This  was  heated  to  50-60®C.  for  5-6  hours  with 
frequent  shaking.  The  solution  was  colored  a dark  blue.  This 
was  due  to  the  formation  of  the  barium  salt.  Carbon  dioxide  was 
then  passed  through  the  warm  solution  and  barium  carbonate  was 
precipitated.  This  was  filtered  off.  The  filtrate  was  evaporated 
under  diminished  pressure  and  a bro'.vn  syrup  was  left. 

This  was  left  to  stand  until  it  was  dry.  ^'^en  viewed  under 
the  microscope  no  crystals  of  alizarin  could  be  seen.  The 
melting  point  was  not  sharp,  but  it  would  start  to  melt  at  about 
85°C.  The  substance  was  washed  with  water  until  it  gave  no 
tests  for  carbohydrates.  Water  was  allowed  to  cstand  upon  it  for 
several  days,  and  still  there  was  no  test  for  carbohydrates,  A 
portion  of  the  substance  was  then  added  to  a dilute  solution  of 

sulphuric  acid  and  hydrolysed  for  several  hours.  The  resulting 


- 15 


solution  gave  positive  tests  for  the  presence  of  carbohydrates 
and  also  showed  free  alizarin,  because  the  latter  was  extracted 
with  ether  and  after  crystalization  gave  the  melting  point  for 
alizarin, 

A mixture  of  emulsin.  and  raaltase  was  added  to  one  gram  of 
the  substance  and  the  mixture  allowed  to  stand  for  several  days 
at  room  temperature.  The  resulting  solution  showed  evidence  of 
hydrolysis  by  enzyme  action  because  positive  tests  were  given 
for  glucose. 

Chloroform,  alcohol,  arnyl  alcohol,  acetone  and  methyl  alco- 
hol were  employed  in  trying  to  cryst^Jize  the  viscous  liquid, 
but  no  positive  results  were  obtained.  Owing  to  the  fact  that 
the  substance  dissolved  slov/ly  in  chloroform  we  tried  to  crystal- 
lize the  substance  from  this  solvent  by  means  of  a vacuum  desib- 
dator.  The  chloroform  solution  containing  the  substance  was 
placed  in  a desiccator.  The  vacuum  pump  was  connected  for  about 
tv/o  minutes.  Then  in  about  an  hour  it  was  again  connected.  This 
was  continued  until  the  solution  had  evaporated.  But  the  sub- 
stance was  again  deposited  as  a viscous  liquid.  The  use  of  this 
solvent  in  a tightly  stoppered  bottle  was  then  resorted  to.  The 
substance  was  heated  in  a small  bottle  until  a portion  had  'dis- 
solved and  a portion  remained  undissolved.  The  bottle  was  tightly 
stoppered  and  set  aside,  nothing  crystalline  formed. 


- 16  - 

In  attempting  a quantitative  hydrolysis,  two  grams  of  the 
acetylated  substance  was  dissolvea  in  a mixture  of  35  cc.  of 
chloroform  and  10  cc.  of  dilute  sulphuric  acid,  and  the  mixture 
hydrolysed  for  several  hours.  The  chloroform  layer  after  wash- 
ing was  then  evaporated,  and  the  resulting  substance  weighed. 

The  result  gave  ,1130  grams,  or  58^  of  the  original  substance. 
Taking  ruberythric  acid  as  a ba^is  in  which  two  molecules  of 
glucose  are  united  with  one  of  alizarin  the  yield  should  be  68'^. 

If  there  was  a union  the  basis  of  conjunction  would  favor  the 
ratio  of  two  to  one  rather  than  one  to  one. 

Owing*  to  the  excess  of  alizarin  which  was  left  each  time 
after  the  extraction  of  the  mixed  mass  of  alizarin  and  aceto- 
bromoglucose,  different  methods  were  tried  in  order  to  increase 
the  amount  of  the  amorphous  substance  which  would  remain.  A 
mixture  of  5 grams  of  acetobromoglucose  and  3,86  grams  of  alizarin 
were  dissolved  in  ten  grams  of  quinoline  and  refluxed  at  170°  for 
one  hour.  The  remaining  quinoline  ?/as  extracted  with  dilute 
sulphuric  acid  and  chloroform.  It  was  evident  that  too  much  heat 
had  been  applied,  because  the  resulting  product  was  partially 
carbonized. 

Five  grams  of  acetobromo  glucose  and  3.66  grams  of  alizarin 
were  intimately  mixed  in  a beaker  and  4 grams  of  quinoline  poured 
over  the  mixture.  3 grams  of  mercury  were  added  as  a catalyst. 

The  mixture  ?/as  heated  on  an  oil  bath  at  110-115*  for  over  and 
hour.  It  was  then  washed  with  100  cc,  of  N sulphuric  acid,  and 
75  cc.  of  chloroform  filtered  and  the  chloroform  extract  evapor- 


- 17 


atsd,  Ths  light  brown  viscous  substance  and  some  free  alizarin 
resulted,  but  the  relative  amount  had  not  increased.  This 
residue  gave  the  same  tests  as  were  applied  to  the  similar  sub- 
stance that  had  been  isolated,  and  gave  the  sane  results. 

A blank  test  was  made  of  menthol  glucoside  which  has  been 

(33) 

successfully  formed  by  this  method.  The  yield  in  this  case  was 
quantitative,  and  the  compound  was  easily  crystallized.  This 
compound  would  not  have  the  complexity  of  a hydroxyanthraquinone 
glucoside,  and  for  this  reason  would  be  more  easily  formed  and 
crystallized. 

B 

Emodin  Glucoside, 

The  procedure  followed  was  similar  to  that  use  in  the  case 
of  alizarin.  Five  grams  of  acetobromoglucose  were  added  to  an 
absolute  alcohol  solution  of  3.3  grains  of  emodin  and  3.06  grams 
of  potassium  hydroxide.  The  mixture  was  allowed  to  stand  for 
a week  in  a tightly  stoppered  flask,  and  then  refluxed  for  an 
hour.  The  deep  red  solution  still  remained.  The  odor  of  ethyl 
acetate  was  evident  when  the  flask  was  opened. 

The  alcoholic  solution  was  diluted  and  exactly  neutralized 
with  hydrochloric  acid.  The  light  brown  precipitate  settled  out. 
This  was  filtered  and  dried  upon  a porous  plats.  After  washing 
with  ether  to  extract  the  free  emodin  and  with  water  to  extract 
any  uncombined  carbohydrate,  the  substance  was  hydrolysed  with 
dilute  sulphuric  acid.  The  resulting  solution  gave  no  test  for 
the  presence  of  a carbohydrate. 


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- 18 


EvTidently  no  combination  had  occurred  between  the  emodin 
and  the  acato-bromoglucose. 

The  use  of  the  method  with  the  shaking  machine  was  then 
resorted  to.  Five  grams  of  acetobromoglucose,  3.3  grams  of  emodin 
2.06  grams  of  potassium  hydroxide  were  added  to  200  cc,  of  water 
and  200  cc.  of  ether.  This  mixture  was  placed  in  the  bottle  which 
was  connected  with  the  shaking  machine.  This  was  run  inter- 
mittently for  twenty-four  hours.  The  ethereal  layer  was  evap- 
orated. The  dark  powder  which  resulted  was  subjected  to  hydroly- 
sis with  dilute  sulphuric  acid,  and  did  not  yield  any  test  for 
the  presence  of  carbohydrates. 

The  dark  powder  gave  no  evidence  of  the  combination  of 
emodin  and  glucose. 

C 

EMODIN  GLUCOSIDE  (Anthra-rhamnoside) 
the  procedure  was  similiar  to  that  carried  out  with  the 
alizarin  and  tetra-acetylbromoglucose,  with  however,  the  use  of 
acetobromorharnnose  in  the  place  of  the  glucose  derivative. 

Five  grams  of  the  acetobromorharnnose  were  added  to  an  ab- 
solute alcohol  solution  of  four  graRis  of  emodin  and  2.48  grams  of 
potassium  hydroxide.  A deep  red  mixture  was  formed.  This  was 
allowed  to  stand  at  room  temperature  for  one  week,  and  then 
refluxed  for  two  hours.  The  odor  of  ethyl  acetate  was  apparent 
when  the  cork  viras  removed.  A dark  powder  had  precipitated  out. 
This  was  filtered  off  and  dried.  It  was  then  washed  with  ether 
and  dried.  It  failed  to  give  a melting  point  belov/  300°C.  and 


- 19 


was  insoluble  in  the  organic  solvents.  Upon  hydrolysis  it  did 

not  yield  any  test  for  carbohydrate.  This  did  not  in  any  way 

(18) 

correspond  to  the  product  which  was  secured  by  Gunton,  but 
upon  analysis  it  was  found  that  the  product  had  been  destroyed 
through  too  much  refluxing. 

The  same  quantities  were  again  mixed  and  added  to  an  ab- 
solute alcohol  solution,  and  permitted  to  stand  for  one  week. 

This  ms  then  refluxed  for  45  minutes.  The  contents  were  poured 
into  a beaker  and  permitted  to  evaporate  at  room  temperature. 

The  resulting  mass  was  a light  brovm  powder.  This  was  dissolved 
in  a small  quantity  of  water  and  exactly  neutralized  with  hydro- 
chloric acid.  A light  brown  precipitate  settled  out.  This  was 
filtered  off,  and  dissolved  in  a mixture  of  alcohol  and  water. 

A slight  portion  did  not  dissolve,  so  this  was  filtered  off.  The 
solution  was  digested  on  the  steam  bath  with  the  occasional 
addition  of  some  distilled  water.  A light  hrorni  powder  soon 
settled  out.  This  was  washed  with  v;ater  and  dried,  then  washed 
in  ether.  The  ether  extract  v;as  evaporated,  and  the  residue  gave 
a melting  point  for  emodin.  After  complete  drying  the  substance 
gave  a melting  point  of  330°,  although  this  was  not  sharp.  A 
portion  of  this  sample  was  intimately  mixed  with  some  natural 
frangulin  and  the  resulting  melting  point  was  315°. 

The  substance  was  wahhed  with  water.  V/hen  the  water  gave 
no  test  for  the  presence  of  carbohydrate^  a sample  was  then 
subjected  to  hydrolysis  with  dilute  sulphuric  acid  for  three 
hours.  The  resulting  solution  gave  positive  tests  for  the 


- 30  - 

presence  of  a carbohydrate,  altho  all  of  the  material  did  not 
hydrolyse  readily.  Extraction  of  the  hydrolysed  mixture  v;ith 
ether  gave  pure  emodin. 

Another  test  was  made  with  the  same  amounts  and  the  same 
procedure  followed.  The  yield  was  the  same  as  in  the  previous 
experiment.  The  amorphous  powders  of  the  two  experiments  were 
mixed  together  and  attempts  were  made  to  secure  a crystalline 
product  through  the  use  of  alcohol,  chloroform  and  acetone.  The 
latter  was  used  because  it  has  been  used  as  a crystallizing 
solvent  for  frangulin.  The  slow  evaporation  method  and  the  use 
of  the  vacuum  were  resorted  to,  but  the  amorphous  powder  was 
always  deposited  when  the  solvent  had  avaporated. 

Although  the  color  of  the  powder, and  melting  point  point 
resembled  frangulin,  the  solubility  of  the  substance  was  not  of 
the  same  class.  Nothing  crystalline  was  obtained.  But  from  the 
properties  found  there  seems  to  have  been  some  combination  of  the 
glucose  derivative  and  the  emodin. 


- 31 


V 

SU1'£MARY 

The  synthesis  of  an  alizarin  gluooside  was  atteii^tsd 
by  means  of  a potassium  salt  method  with  an  acetyl 
derivative  of  glucose.  Nothing  of  a glucosidic  compound 
ms  isolated  from  the  result.  By  using  the  saurjematerial 
but  by  trying  to  effect  the  combination  through  shaking 
in  an  etheral  solution  another  trial  was  made.  This 
failed  to  yield  any  glucoside. 

The  formation  of  a substance  by  heating  the  glucose 
derivative  and  alizarin  in  quinoline  gave  tests  that  a com- 
bination had  taken  place  in  the  ratio  of  two  molecules  of 
sugar  to  one  of  alizarin. 

By  means  of  the  potassium  salt  method  v;ith  emodin  and 
rhamnose  a compound  v/as  formed  which  had  similar  melting 
point  and  color  to  the  natural  oc curing  frangulin,  but  its 
solubility  differed  from  the  latter. 


- 33- 


VI 

BIBLIOGRAPHY 

(1)  Robiqust  and  Colin  - Annanles  de  Chemie  (1837),  335 

(2)  Decaisne  - J.  prakt.  Chem,  (1838),  1^,  393 

(3)  Rochledsr  - Ann.  Chem.  346;  _80,  321;  0^,  205. 

(4)  Schunk  - J.  Chsm.  Soc.  (1860),  12,  316 

(5)  Stokes  - J.  Chem.  Soc.  (i860),  13,  319 

(6)  Schunk  and  Harchlewski  - J.  Chem.  Soc  (1893),  _63,  969 

(7)  Lieberrnann  and  Bergami  - Ber.  Chem.  Ges.(l887),  2241 

(8)  Perkin  - J.  Chem.  Soc.  (1897),  H94 

(9)  Ivliiller  and  la  Rue  - Jahresber,  (1857)  517. 

(10)  Muller  - J.  Chem.  Soc.  (1911),  967. 

(11)  Tutin  and  Clewer  - J,  Chem.  Soc.  ('1911  )_^,  I,  946. 

(13)  Jowett  and  Potter,  J.  Chem,  Soc.  _81,  1575 

(13)  Tutin  - J.  Chem.  Soc.  (1913),  103,  2006 

(14)  Bailsy  - J.  Ind.  and  Eng.  Chem.  (1914)  ,_6,  320 

(15)  Binswanger  - Ann.  de  Chem.  u.  Phar.  23.*  ^56. 

(16)  Faust  - Ann.  de  Chem.  u.  Phar.  165,  339 
Kubly 

(17) .APharm.  Zeitsch.  F.  Russl,  V,  part.  3. 

Gunton 

(18) ^hesis,  University  of  111.  (1931) 

0 

(19)  Colley  Comp.  rend.  70,  401 

(30)  Michael  - Ber.  d.  deut.  Chem.  Ges,  (1881),  lA,  3097 

(21)  Schiff  - Ann.  144,  19  (1886) 

(22)  Druin  - Bull.  Soc.  Chira.  (ill),  13,  5.  (1895) 

(33)  Irvine  - J.  Chem.  Soc.  (1913)  103 . 41. 


- 33 


(34) 

Ryan  - 

Proc.  Roy^  Irish  Acad.  (1903),  34,  379 

(35) 

Ivlauthner 

- 2T.  prakt.  Chem.  (1910),  _83,  271 

(36) 

Sal  way 

- J.  Chem.  Soc.  (1913),  lOZ,  1033 

(37) 

Schneider  - Ber.  (1914)  1356_1369;  3318-3334 

(38) 

Fischer 

- Ber. (1893) ,16,  3938 
Ber.  (1895)  ,38,  1145 
Annalen . (1911 ) , 383,  68 

(39) 

Walton 

- J.A.C.S.  (1931)  ,43,137 

(30) 

Thorpe, 

Diet.  Applied  Chem.  Vol.  I,  79. 

(31) 

Dale, 

J.  A.C.S,  (1916)  38,  3187 

(33) 

Fischer 

- Ber.  4_5,  3468  (1913) 

(33) 

Fischer 

- Ber.  (1917),  50,  711 

